Methods of synthesizing 2-[4-[(2,3,4-trimethoxyphenyl)methyl]piperazin-1-yl]ethyl pyridine-3-carboxylate

ABSTRACT

The invention provides methods of chemical synthesis of the pharmacological agent 2-[4-[(2,3,4-trimethoxyphenyl)methyl]piperazin-1-yl]ethyl pyridine-3-carboxylate, also called CV-8972. The methods entail formation of a free base form of 2-[4-[(2,3,4-trimethoxyphenyl)methyl]piperazin-1-yl]ethanol, also called CV-8814, as intermediate without producing a salt form of CV-8814.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of, and priority to, U.S.Provisional Patent Application No. 63/046,123, filed Jun. 30, 2020, thecontents of which are incorporated by reference.

FIELD OF THE INVENTION

The invention relates to methods of chemical synthesis of thepharmacological agent2-[4-[(2,3,4-trimethoxyphenyl)methyl]piperazin-1-yl]ethylpyridine-3-carboxylate.

BACKGROUND

Heart disease is the leading cause of death worldwide, accounting for 15million deaths across the globe in 2015. In many forms of heart disease,decreased cardiac efficiency stems from changes in mitochondrial energymetabolism. Mitochondria are sub-cellular compartments in whichmetabolites derived from glucose and fatty acids are oxidized to producehigh-energy molecules. Increasing fatty acid oxidation in the heartdecreases glucose oxidation, and vice versa. Glucose oxidation is a moreefficient source of energy, but in certain types of heart disease, suchas angina, heart failure, ischemic heart disease, and diabeticcardiomyopathies, fatty acid oxidation predominates in cardiacmitochondria. As a result, the pumping capacity of the heart is reduced.

SUMMARY

CV-8972, which has the IUPAC name2-[4-[(2,3,4-trimethoxyphenyl)methyl]piperazin-1-yl]ethylpyridine-3-carboxylate and the following structure:

was recently identified as a promising therapeutic candidate forcardiovascular conditions. Prior schemes for synthesis of CV-8972require formation of a free base form of2-[4-[(2,3,4-trimethoxyphenyl)methyl]piperazin-1-yl]ethanol, also calledCV-8814, and conversion of the free base form of CV-8814 to ahydrochloride salt. In such schemes, CV-8814 must then be converted backto its free base form for coupling to nicotinic acid to form the freebase form of CV-8972.

The invention provides a CV-8972 synthesis scheme that bypasses thereversible conversion of CV-8814 between the free base and HCl saltforms. In the schemes provided herein, the free base form of CV-8814 isformed in a reductive amination reaction, and the free base product isused directly as a substrate for coupling to nicotinic acid to formCV-8972. Because fewer steps are required, the synthesis schemes of theinvention are simpler, faster, and provide better yields than priormethods of making CV-8972.

In an aspect, the invention provides methods for preparing a compound ofFormula (X):

by performing the steps of:

reacting 2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-olto produce a free base form of a compound of Formula (IX):

and

reacting the free base form of the compound of Formula (IX) withnicotinic acid to produce the compound of Formula (X), wherein themethod does not comprise producing a salt form of the compound ofFormula (IX).

The step of reacting 2,3,4-trimethoxybenzaldehyde and2-(piperazin-1-yl)ethan-1-ol may include one or more solvents,catalysts, or other chemicals. The step of reacting2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-ol mayinclude one or more of sodium triacetoxyborohydride, acetic acid, and2-methyltetrahydrofuran.

The step of reacting 2,3,4-trimethoxybenzaldehyde and2-(piperazin-1-yl)ethan-1-ol may be performed at a defined temperature.The step of reacting 2,3,4-trimethoxybenzaldehyde and2-(piperazin-1-yl)ethan-1-ol may be performed at from about 10° C. toabout 30° C., from about 15° C. to about 30° C., from about 20° C. toabout 30° C., from about 25° C. to about 30° C., from about 10° C. toabout 25° C., from about 15° C. to about 25° C., from about 20° C. toabout 25° C., from about 10° C. to about 20° C., or from about 15° C. toabout 20° C.

The step of reacting 2,3,4-trimethoxybenzaldehyde and2-(piperazin-1-yl)ethan-1-ol may not include a specific solvent,catalyst, or other chemical. The step of reacting2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-ol may notinclude dichloromethane.

The step of reacting the free base form of the compound of Formula (IX)with nicotinic acid may produce a free base form of the compound ofFormula (X).

The step of reacting the free base form of the compound of Formula (IX)with nicotinic acid may include one or more solvents, catalysts, orother chemicals. The step of reacting the free base form of the compoundof Formula (IX) with nicotinic acid may include one or more of1-ethyl-3-(3-dimethylaminopropyl)carbodiimide,4-(dimethylamino)pyridine, and dichloromethane.

The step of reacting the free base form of the compound of Formula (IX)with nicotinic acid may be performed at a defined temperature. The stepof reacting the free base form of the compound of Formula (IX) withnicotinic acid may be performed at from about 15° C. to about 30° C.,from about 20° C. to about 30° C., from about 25° C. to about 30° C.,from about 15° C. to about 25° C., from about 20° C. to about 25° C., orfrom about 15° C. to about 20° C.

The method may include converting the free base form of the compound ofFormula (X) to a salt form of the compound of Formula (X). The salt formof the compound of Formula (X) may be a HCl salt. The salt form of thecompound of Formula (X) may be monohydrate.

The step of converting the free base form of the compound of Formula (X)to the salt form of the compound of Formula (X) may include one or moresolvents, catalysts, or other chemicals. The step of converting the freebase form of the compound of Formula (X) to the salt form of thecompound of Formula (X) may include one or more of HCl and methyl ethylketone.

The step of converting the free base form of the compound of Formula (X)to the salt form of the compound of Formula (X) may be performed at adefined temperature. The step of converting the free base form of thecompound of Formula (X) to the salt form of the compound of Formula (X)may be performed at from about 40° C. to about 60° C., from about 45° C.to about 60° C., from about 50° C. to about 60° C., from about 55° C. toabout 60° C., from about 40° C. to about 55° C., from about 45° C. toabout 55° C., from about 50° C. to about 55° C., from about 40° C. toabout 50° C., from about 45° C. to about 50° C., from about 40° C. toabout 50° C., about 40° C., about 45° C., about 50° C., about 55° C., orabout 60° C.

The method may include converting the salt form of the compound ofFormula (X) from a first crystal form to a second crystal form. Each ofthe first and second crystal forms may independently be Form A, Form B,Form C, Form D, or Form E.

The step of converting the salt form of the compound of Formula (X) froma first crystal form to a second crystal form may include one or more ofchanging the solvent of the salt form of the compound of Formula (X) andincubating the salt form of the compound of Formula (X), at about 60° C.

The method may be performed without the use of one or more solvents,catalysts, or other chemicals. The method may be performed without theuse of one or more of dioxane, ethylacetate, or potassium carbonate.

The method may include purifying the free base form of the compound ofFormula (IX). The method may include crystallizing the free base form ofthe compound of Formula (IX).

In another aspect, the invention provides methods of preparing acompound of Formula (X) by performing the steps of:

reacting a compound of Formula (1):

with a compound of Formula (2):

to produce a free base form of a compound of Formula (IX):

reacting the free base form of a compound of Formula (IX) with acompound of Formula (3):

to produce a free base form of the compound of Formula (X); and

converting the free base form of the compound of Formula (X) to a HClsalt of the compound of Formula (X),

wherein the method does not comprise producing a salt form of thecompound of Formula (IX).

The method may include purifying the free base form of the compound ofFormula (IX). The method may include crystallizing the free base form ofthe compound of Formula (IX).

DETAILED DESCRIPTION

The recently-identified compound CV-8972 holds promise as a therapeuticagent for treating a variety of conditions, including cardiovascularconditions, rheumatic diseases, fibrosis, and cancer. CV-8972, which hasthe IUPAC name 2-[4-[(2,3,4-trimethoxyphenyl)methyl]piperazin-1-yl]ethylpyridine-3-carboxylate and the following structure:

is metabolized in the body into two sets of products that increasemitochondrial energy production in different ways. In an initialreaction, the molecule is split into CV-8814, which has the followingstructure:

and nicotinic acid. Over time, CV-8814 converted in the body totrimetazidine. Both CV-8814 and trimetazidine inhibit beta-oxidation offatty acids and therefore shift mitochondrial metabolism towardoxidation of glucose, a more oxygen-efficient source of energy.Nicotinic acid serves as precursor for synthesis of nicotinamide adeninedinucleotide (NAD⁺). NAD⁺ promotes mitochondrial respiration to driveATP synthesis, regardless of whether glucose or fatty acids are used asthe carbon source. Thus, the two sets of products that result frombreakdown of CV-8972 in vivo act synergistically to stimulate energyproduction in mitochondria in cardiac tissue and other cell types.CV-8972 and its mechanism of action are described in U.S. Pat. No.10,556,013, the contents of which are incorporated herein by reference.

U.S. Pat. No. 10,556,013 also provides a scheme for synthesis ofCV-8972. The scheme entails formation of a free base form of CV-8814 byreductive amination of 2,3,4-trimethoxybenzaldehyde and2-(piperazin-1-yl)ethan-1-ol. Due to the difficulty of isolating CV-8814in a solid form in this prior method, the product of this reaction isthen converted to a hydrochloride salt of CV-8814. However, CV-8814 mustbe converted back to its free base form for use in the esterificationreaction with nicotinic acid that produces CV-8972.

The invention provides CV-8972 synthesis schemes in which the free baseform of CV-8814 formed as a product in the reductive amination reactioncan be used directly as a substrate in the esterification reaction. Theinvention is based in part on the identification of conditions thatimprove the stability of CV-8814 free base and allow the free base formto be crystallized. Thus, the schemes provided herein obviate the needto convert CV-8814 from its free base form to a HCl salt and then backto the free base form. Consequently, the invention provides simpler,quicker, and higher-yield methods for making CV-8972.

The invention provides methods for preparing a compound of Formula (X):

by performing the steps of:

reacting 2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-olto produce a free base form of a compound of Formula (IX):

and

reacting the free base form of the compound of Formula (IX) withnicotinic acid to produce the compound of Formula (X),

wherein the method does not comprise producing a salt form of thecompound of Formula (IX).

2,3,4-trimethoxybenzaldehyde has the following structure:

2-(piperazin-1-yl)ethan-1-ol has the following structure:

Nicotinic acid has the following structure:

The step of reacting 2,3,4-trimethoxybenzaldehyde and2-(piperazin-1-yl)ethan-1-ol may include one or more solvents,catalysts, or other chemicals. The step of reacting2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-ol mayinclude one or more of sodium triacetoxyborohydride, acetic acid, and2-methyltetrahydrofuran.

The step of reacting 2,3,4-trimethoxybenzaldehyde and2-(piperazin-1-yl)ethan-1-ol may be performed at a defined temperature.The step of reacting 2,3,4-trimethoxybenzaldehyde and2-(piperazin-1-yl)ethan-1-ol may be performed at from about 10° C. toabout 30° C., from about 15° C. to about 30° C., from about 20° C. toabout 30° C., from about 25° C. to about 30° C., from about 10° C. toabout 25° C., from about 15° C. to about 25° C., from about 20° C. toabout 25° C., from about 10° C. to about 20° C., or from about 15° C. toabout 20° C.

The step of reacting 2,3,4-trimethoxybenzaldehyde and2-(piperazin-1-yl)ethan-1-ol may not include a specific solvent,catalyst, or other chemical. The step of reacting2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-ol may notinclude dichloromethane.

The step of reacting the free base form of the compound of Formula (IX)with nicotinic acid may produce a free base form of the compound ofFormula (X).

The step of reacting the free base form of the compound of Formula (IX)with nicotinic acid may include one or more solvents, catalysts, orother chemicals. The step of reacting the free base form of the compoundof Formula (IX) with nicotinic acid may include one or more of1-ethyl-3-(3-dimethylaminopropyl)carbodiimide,4-(dimethylamino)pyridine, and dichloromethane.

The step of reacting the free base form of the compound of Formula (IX)with nicotinic acid may be performed at a defined temperature. The stepof reacting the free base form of the compound of Formula (IX) withnicotinic acid may be performed at from about 15° C. to about 30° C.,from about 20° C. to about 30° C., from about 25° C. to about 30° C.,from about 15° C. to about 25° C., from about 20° C. to about 25° C., orfrom about 15° C. to about 20° C.

The method may include converting the free base form of the compound ofFormula (X) to a salt form of the compound of Formula (X). The salt formof the compound of Formula (X) may be a HCl salt. The salt form of thecompound of Formula (X) may be monohydrate.

The step of converting the free base form of the compound of Formula (X)to the salt form of the compound of Formula (X) may include one or moresolvents, catalysts, or other chemicals. The step of converting the freebase form of the compound of Formula (X) to the salt form of thecompound of Formula (X) may include one or more of HCl and methyl ethylketone.

The step of converting the free base form of the compound of Formula (X)to the salt form of the compound of Formula (X) may be performed at adefined temperature. The step of converting the free base form of thecompound of Formula (X) to the salt form of the compound of Formula (X)may be performed at from about 40° C. to about 60° C., from about 45° C.to about 60° C., from about 50° C. to about 60° C., from about 55° C. toabout 60° C., from about 40° C. to about 55° C., from about 45° C. toabout 55° C., from about 50° C. to about 55° C., from about 40° C. toabout 50° C., from about 45° C. to about 50° C., from about 40° C. toabout 50° C., about 40° C., about 45° C., about 50° C., about 55° C., orabout 60° C.

The compound of Formula (X) may exist in at least five crystal forms:Form A, Form B, Form C, Form D, and Form E. Form A is monohydrate, andForms B, D, and E are anhydrous. The method may include converting thecompound of Formula (X) from a first crystal form to a second crystalform. Each of the first and second crystal forms may independently beForm A, Form B, Form C, Form D, or Form E. The method may include one ormore of the following conversions of the compound of Formula (X): froman anhydrous form to a hydrated form; from a hydrated form to ananhydrous form; from one anhydrous form to another; and from onehydrated form to another.

The step of converting the salt form of the compound of Formula (X) froma first crystal form to a second crystal form may include one or more ofchanging the solvent of the salt form of the compound of Formula (X) andincubating the salt form of the compound of Formula (X), at about 60° C.

The method may be performed without the use of one or more solvents,catalysts, or other chemicals. The method may be performed without theuse of one or more of dioxane, ethylacetate, or potassium carbonate.

The method may include purifying the free base form of the compound ofFormula (IX). The method may include crystallizing the free base form ofthe compound of Formula (IX).

EXAMPLES Example 1

Introduction

CV-8972 was synthesized according to Scheme 1.

Step 1 is a reductive amination using 2,3,4-trimethoxybenzaldehyde and2-(piperazin-1-yl)ethan-1-ol starting materials, with sodiumtriacetoxyborohydride (STAB) as the reductant, in the presence ofcatalytic acetic acid (AcOH), and 2-methyltetrahydrofuran (2-MeTHF) assolvent. After the reaction is completed, an aqueous workup, solventexchange to MTBE, and recrystallization from MTBE/n-heptane forms theintermediate CV-8814 Free Base (CV8814 Free Base).

In step 2, CV-8814 Free Base (CV8814 Free Base) undergoes acid couplingwith nicotinic acid in the presence of1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) and catalytic4-(dimethylamino)pyridine (DMAP) in dichloromethane (DCM) solvent. Afteraqueous workup, CV8972 Free Base is formed. Solvent exchange to2-butanone (MEK) followed by addition into concentrated HCl in MEK formsCV8972 Monohydrate intermediate.

The final step 3 is a form conversion in a mixture of water, methanol,and MEK at 60° C.±5° C. followed by a precipitation by the addition ofMEK to obtain the desired form A of final product CV-8972 by XRPDanalysis.

Manufacturing Details

Manufacturing details are provided in Table 1.

TABLE 1 Amount Amount Started Produced Yield HPLC Step# BOP# (Kg) (Kg)(%) Purity (%) 1 2493-1903-00484 27.5 kg 34.0 kg 76.0% 100.0% 22479-1903-00489 28.8 kg 48.6 kg 96.4%  99.1% 3 2479-1904-00494 48.5 kg41.7 kg 86.0%  99.9%

Production Details

Step 1, formation of CV8814 Free Base (2493-1903-00487), was performedaccording to Scheme 2.

Production details for step 1 are provided in Table 2.

TABLE 2 Actual Equiv. Expected Quantity Identity Vendor/Grade/ MW Moles/Moles Quantity Used Name Catalog # g/mol mol-LR moles Units Units2,3,4-Trimethoxy- Oakwood/98%/ 196.20 1.00 140.16  27.5 kg  27.5 kgbenzaldehyde 078721; Chem-Impex/   NA/26868; Chem-   Impex/NA/26868  1-(2-hydroxyethyl)piperazine Aldrich/98%/ 130.19 1.1  154.18  20.1 kg 20.0 kg H28807 2-Methyltetrahydrofuran (2- Aldrich/≥99.5%/ N/A N/A N/A309.5 kg  338.3 kg MeTHF) 155810 Plus As Needed Acetic Acid (AcOH)Aldrich/≥99%/ N/A N/A N/A  1.40 kg 1.4101 kg A6283 Sodium TriacetoxyOakwood/95%/ 211.94 2.00 280.32  59.4 kg Plus  60.3 kg Borohydride(STAB) 044864 As Needed Sodium Hydroxide solution, Aldrich/50% in H₂O/40 6.09 853.57  68.3 kg  68.3 kg 50% in H₂O (50% NaOH in 415413 H₂O)Sodium Chloride Aldrich/ACS/ N/A N/A N/A  19.0 kg  19.0 kg (NaCl) S9888Methyl tert-Butyl Ether Aldrich/ACS/ N/A N/A N/A 417.3 Plus  552.1 kg(MTBE) 443808; Oakwood/ As Needed ACS/099538 Water Mediatech/WFI/25- N/AN/A N/A 323.4 kg  323.4 kg 055-X¹; RMBI/USP/ WPW-USN-2XL HeptaneAldrich/99%/ N/A N/A N/A 159.8 kg  162.1 kg H2198; Oakwood/ ACS/044743;BDH/ ACS/BDH1127 Nitrogen Gas Airgas ≥ N/A N/A N/A Quantity Quantity(house system) 99% Sufficient Sufficient (QS)

-   1) Sodium Triacetoxy Borohydride (STAB; 60.3 kg; CHP Lot #:    181-190220) and 2-MeTHF (189.2 kg; CHP Lot #: 234-190227) was    charged to reactor R-401.-   2) The contents of R-401 were agitated and the temperature was    adjusted to 15° C.±5° C.-   3) 2,3,4-trimethoxybenzaldehyde (27.5 kg; CHP Lot #: 275-190311,    142-190212) and 2-MeTHF (71.1 kg: CHP Lot #: 234-190227) was charged    to reactor R-402.-   4) Agitation of reactor R-402 was started.-   5) 2-(piperazin-1-yl)ethan-1-ol (20.0 kg; CHP Lot #: 173-190220) and    2-MeTHF (47.3 kg; CHP Lot #: 234-190227) was charged to reactor    R-402.-   6) The contents of reactor R-402 were agitated for at least 5    minutes.-   7) Acetic Acid (1.41 kg: CHP Lot #: 151-190214) was charged to    reactor R-401 while keeping the temperature of the mixture below 25°    C.-   8) T_(max)=14.4° C.-   9) The contents of reactor R-402 were transferred over 1 h 19 min    while keeping the temperature of the mixture below 25° C.-   10) T_(max)=29.2° C. (the temperature went out of range during the    addition)-   11) The reaction was approved to proceed forward-   12) The temperature of the contents of R-401 were adjusted to 20°    C.±5° C. and agitated for at least 6 hours at 20° C.±5° C.-   13) The contents of R-401 were sampled after approximately 19 hours.-   14) Analysis of the sample by QC indicated no peak of    2,3,4-trimethoxybenzaldehyde was detected. (specification ≤1.5 area    %)-   15) The temperature of the contents of R-401 were adjusted to 15°    C.±5° C.-   16) Water (247.5 kg; CHP Lot #: 232-190227) was charged to R-401    while keeping the temperature below 25° C.-   17) T_(max)=14.6° C.-   18) The temperature of the contents of R-401 was adjusted to 20°    C.±5° C. and the contents were agitated for 30 minutes.-   19) The contents of R-401 were allowed to settle for 30 minutes.-   20) Phase cut was performed with the aqueous layer being transferred    to R-402.-   21) MTBE (203.8 kg; CHP Lot #: 207-190222) was charged to R-402.-   22) The temperature of the contents of R-402 was adjusted to 0°    C.±5° C.-   23) 50% NaOH (68.3 kg; CHP Lot #: 145-190213, 150-190214) was    charged to R-402 while maintaining a temperature below 25° C.-   24) T_(max)=11.5° C.-   25) After complete addition, the temperature of the contents of    R-402 was adjusted to 20° C.±5° C.-   26) The contents in R-402 were agitated for at least 30 min,    followed by allowing the contents to settle for at least 30 min.-   27) A phase cut was performed with the aqueous layer being    transferred to reactor R-401 and the organic layer remaining in    reactor R-402.-   28) MTBE (61.0 kg; CHP Lot #: 207-190222) was charged to R-401.-   29) The temperature of the contents of R-401 was adjusted to 20°    C.±5° C.-   30) The contents in R-401 were agitated for at least 15 min,    followed by allowing the contents to settle for at least 15 min.-   31) A phase cut was performed with the aqueous layer being    transferred to a drum and the organic later remaining in reactor    R-401.-   32) FIO pH check of drummed aqueous layer pH=13.20-   33) The contents in R-402 were transferred to R-401.-   34) 20% NaCl solution (94.4 kg; NaCl: 19.0 kg, CHP Lot #:    156-190214; Water (75.9 kg; CHP Lot #: 232-190227) was charged to    R-401, agitated for at least 15 minutes and allowed to settle for at    least 30 minutes.-   35) A phase cut was performed with the aqueous layer being    transferred to a drum.-   36) The solution in R-401 was distilled under reduced pressure while    maintaining a temperature <45° C. to approximately ˜55 L total    volume.-   37) MTBE (61.1 kg; CHP Lot #: 207-190222) was charged to R-401.-   38) The contents in R-401 were distilled under reduced pressure    while maintaining a temperature <45° C. to approximately ˜82 L total    volume.-   39) MTBE (61.1 kg; CHP Lot #: 207-190222) was charged to R-401.-   40) The contents in R-101 were distilled under reduced pressure    while maintaining a temperature <45° C. to approximately ˜82 L total    volume.-   41) The contents of R-401 were sampled (IPC sample:    2493-1903-00484-85-01) to check the water content of the solution by    KF analysis.-   42) KF=1.6% (specification ≤0.5%)-   43) MTBE (61.1 kg; CHP Lot #: 207-190222) was charged to R-401.-   44) The contents in R-101 were distilled under reduced pressure    while maintaining a temperature <45° C. to approximately ˜82 L total    volume.-   45) The contents of R-401 were sampled (IPC sample:    2493-1903-00484-88-01) to check the water content of the solution by    KF analysis.-   46) KF=0.8% (specification ≤0.5%)-   47) MTBE (61.1 kg; CHP Lot #: 207-190222) was charged to R-401.-   48) The contents in R-101 were distilled under reduced pressure    while maintaining a temperature <45° C. to approximately ˜82 L total    volume.-   49) The contents of R-401 were sampled (IPC sample:    2493-1903-00484-91-01) to check the water content of the solution by    KF analysis.-   50) KF=0.4% (specification ≤0.5%)-   51) MTBE (30.9 kg; CHP Lot #: 207-190222) was charged to R-401.-   52) The temperature of the contents of R-401 was adjusted to 40°    C.±5° C.-   53) Heptane (56.3 kg; CHP Lot #: 233-190227) was charged over 8 min    to R-401 while maintaining the temperature at 40° C.±5° C.-   54) T_(min)=38.1° C.-   55) A FIO sample (FIO sample: 2493-1903-00484-100-01) was taken to    observe the ratio of MTBE:Heptane of the contents in R-401. The    ratio was 1.5:6.0.-   56) The temperature of the contents of R-401 were adjusted to 28°    C.±5° C. (Target 26° C. to 29° C.). over at least 30 min and    agitated at that temperature for at least 30 min.-   57) Solid formation was observed.-   58) The temperature of the contents of R-401 were adjusted to 30°    C.±3° C. and agitated for at least 30 min.-   59) Heptane (56.4 kg; CHP Lot #: 233-190227) was charged over 24 min    to R-401 while maintaining the temperature at 30° C.±5° C.-   60) T_(min)=30.2° C.-   61) The temperature of the contents of R-401 were adjusted to 30°    C.±3° C. and agitated for at least 20 min.-   62) The temperature of the contents of R-401 were adjusted to 20°    C.±5° C. over at least 30 min and agitated for at least 20 min.-   63) The temperature of the contents of R-401 were adjusted to 5°    C.±5° C. over at least 30 min and agitated for at least 30 min.-   64) The solid was collected on Filter-FD-400.-   65) The contents of Filter-FD-400 were washed with cold heptane    (49.4 kg; CHP Lot #: 233-190227).-   66) The contents of Filter-FD-400 were dried under vacuum at ≤25° C.    with a stream of N2 for at least 16 hours.-   67) An IPC sample (IPC sample: 2493-1903-00484-122-01) was submitted    to QC for LOD.    -   LOD=0.20% (specification ≤0.5%)-   68) The product CV-8814 Free Base (CV8814 Free Base) was double    bagged, goose necked, and weighed.-   69) Analysis of dried material (IPC sample: 2493-1903-00484-122-01):    -   Appearance: White to off white solid    -   Weight: 34.0 kg (78.0% yield)    -   HPLC Purity=100.0%    -   1H NMR: conforms to structure-   70) A 5 kg portion of CV-8814 Free Base (CV8814 Free Base) was    removed from the bulk material double bagged, goose necked, and set    aside for release under the Lot #2493-1903-00484.

Step 2, formation of CV8972 Monohydrate (2479-1903-00489), was performedaccording to Scheme 3.

Production details for steps 2a and 2b are provided in Table 3.

TABLE 3 Actual Equiv. Expected Quantity Identity Vendor/Grade/ MW Moles/Moles Quantity Used Name Catalog # g/mol mol-LR moles Units Units CV8814Free Base (IMB- CHP/NA/CHP Lot 310.39 1.00 92.79  28.8 kg  28.8 kg1028814) # 2493-1903-00484 Nicotinic Acid Aldrich/≥98%/ 123.11 1.5139.19  17.1 kg  17.1 kg N4126; Alfa Aesar/ 99%/A12683 CV8814 Free Base(IMB- CHP/NA/CHP Lot 310.39 1.00 92.79  28.8 kg  28.8 kg 1028814) #2493-1903-00484 Nicotinic Acid Aldrich/≥98%/ 123.11 1.5 139.19  17.1 kg 17.1 kg N4126; Alfa Aesar/ 99%/A12683 Dichloromethane (DCM)Aldrich/ACS/ N/A N/A N/A 536.2 kg 533.4 kg D65100 EDC Oakwood/99%/191.71 1.5 139.19  26.7 kg  26.7 kg 024810; Chem-Impex/ ≥99%/000504-(Dimethylamino)pyridine Aldrich/≥99%/ 122.17 0.15 13.92  1.70 kg  1.70kg (DMAP) 107700 Hydrochloric Acid (Conc Avantor/ACS/ 36.46 3.3 306.21 30.2 kg  30.2 kg HCl) 2612-X¹ Sodium Bicarbonate Aldrich/ACS/ N/A N/AN/A   4.0 kg  4.00 kg (NaHCO₃) S6014 Water Mediatech/WFI/ N/A N/A N/A140.9 kg 140.4 kg 25-055-X¹; RMBI/ USP/ WPW-USN-2XL Methyl Ethyl KetoneOakwood/99.9%/ N/A N/A N/A 858.0 kg 857.8 kg (MEK) 075238 Nitrogen GasAirgas ≥ N/A N/A N/A Quantity Quantity (house system) 99% SufficientSufficient (QS)

-   1) Nicotinic Acid (17.1 kg, CHP Lot #201-190222) and DCM (153.0 kg,    CHP Lot #328-190326) were charged to reactor R-401.-   2) The contents of R-401 was agitated and the temperature was    adjusted to 15±5° C.-   3) CV8814 Free Base (28.8 kg, CHP Lot #2493-1903-00484), EDC (26.7    kg, CHP Lot #147-190213), DMAP (1.70 kg, CHP Lot #152-190214), and    DCM (306.6 kg, CHP Lot #328-190326) were charged to reactor R-402.-   4) The contents of the R-402 were agitated for at least 20 minutes.-   5) The contents of R-402 were transferred to R-401 over at least 30    minutes while keeping the temperature below 25° C.-   6) T_(max)=20.0° C.-   7) The temperature of the contents of R-401 were adjusted to    20±5° C. and was agitated for at least 16 hours.-   8) The contents of R-401 were sampled after approximately 16 hours.-   9) Analysis of the sample by QC indicated 0% (0.05%) of CV-8814 Free    Base (CV8814 Free Base) with respect to CV8972 was detected    (Specification: ≤1% CV8814 Free Base).-   10) The contents of the R-401 were adjusted to 10±5° C.-   11) Water (29.2 kg, CHP Lot #329-190326) was slowly added while    keeping the temperature below 25° C.-   12) T_(max)=12.3° C.-   13) The temperature of the contents of R-401 was adjusted to 20±5°    C., agitated for at least 15 min and allowed to settle for at least    15 min.-   14) Phases were separated.-   15) The lower organic layer containing product was transferred to    reactor R-402. The aqueous layer was sent to a drum.-   16) Water (29.0 kg, CHP Lot #329-190326) was charged to the reactor.-   17) The biphasic mixture was agitated for 15 min and allowed to    settle for 15 min.-   18) Phases were separated.-   19) The organic layer containing product was transferred to reactor    R-401. The aqueous layer was sent to a drum.-   20) 8% NaHCO₃ aqueous solution (Sodium Bicarbonate, 4.0 kg, CHP Lot    #192-190221; Water, 53.2 kg, CHP Lot #329-190326) was added to the    reactor.-   21) The mixture was agitated for at least 15 min and allowed to    settle for at least 15 min.-   22) Phases were separated.-   23) The lower organic layer containing product was transferred to    reactor R-402. The aqueous layer was sent to a drum.-   24) Water (29.0 kg, CHP Lot #329-190326) was added to the reactor.-   25) The mixture was agitated for at least 15 min and allowed to    settle for at least 15 min.-   26) R-401 was cleaned with Water (17.6 kg, CHP Lot #329-190326) and    MEK (5.9 kg, CHP Lot #330-190326) and dried with a stream of N2.-   27) Phases were separated.-   28) The lower organic layer containing product was transferred to    reactor R-401. The aqueous layer was sent to a drum.-   29) The contents of R-401 were concentrated under reduced pressure    to approx. 72 L keeping the temperature below 45° C.-   30) T_(max)=32.0° C.-   31) MEK (139.0 kg, CHP Lot #330-190326) was charged to R-401.-   32) The contents of R-401 were concentrated under reduced pressure    to approx. 72 L keeping the temperature below 45° C.-   33) T_(max)=32.0° C.-   34) MEK (139.1 kg, CHP Lot #330-190326) was charged to R-401.-   35) The contents of R-401 were concentrated under reduced pressure    to approx. 72 L keeping the temperature below 45° C.-   36) T_(max)=29.2° C.-   37) FIO 1H NMR to determine the DCM:MEK ratio was taken.    DCM:MEK=1:214.9-   38) MEK (185.5 kg, CHP Lot #330-190326) was charged to R-401.-   39) MEK (208.7 kg, CHP Lot #330-190326) and conc. HCl (30.2 kg, CHP    Lot #274-190311) were charged to a cleaned R-402.-   40) The temperature of the contents of reactor was adjusted to 25±5°    C.-   41) The contents of R-401 were transferred to R-402 over approx. 1    hour while maintaining a temperature below 35° C.-   42) T_(max)=27.2° C.-   43) The temperature of the contents of R-402 was heated to 50±5° C.    and was agitated for at least 1 hour.-   44) The temperature of the contents of the reactor was cooled to    20±5° C. over 2 hours.-   45) The contents of the reactor were agitated at 20±5° C. for 15    hours.-   46) The solid was filtered.-   47) The filter cake was rinsed with MEK (58.0 kg, CHP Lot    #330-190326).-   48) The filter cake was rinsed with MEK (58.0 kg, CHP Lot    #330-190326).-   49) The wet cake was dried in tray dryer 20-25° C. without nitrogen    bleed for at least 16 hours.-   50) An IPC sample (IPC sample: 2479-1903-00489-87-01) was submitted    to QC for KF.    -   Water content (Specification: ≤4% by cKF): 3.3%-   51) The product CV8972 Monohydrate was doubled bagged, goose necked,    and weighed.-   52) Analysis of the dried material (IPC sample:    2479-1903-00489-87-01): Appearance: White to off-white solid    -   Weight: 48.6 kg (96.4% yield)    -   1H NMR: conforms to structure    -   HPLC purity (area %): 99.1%    -   FIO Residual Solvents GC Analysis:        -   2-MeTHF: No Peak        -   DCM: No Peak        -   MTBE: No Peak        -   Heptane: No Peak        -   MEK: 343 ppm        -   Acetic Acid: 874 ppm

Step 3, formation of CV-8972 (2479-1904-00494), was performed accordingto Scheme 4.

Production details for step 3 are provided in Table 4.

TABLE 4 Actual Equiv. Expected Quantity Identity Vendor/Grade/ MW Moles/Moles Quantity Used Name Catalog # g/mol mol-LR moles Units Units CV8972CHP/NA/CHP Lot #2479- 542.88 1 89.34   48.5 kg   48.5 kg monohydrate1903-00489 Methanol Aldrich/ACS/179337; N/A N/A N/A  230.5 kg  230.5 kgPharmco/WORLD/ 339WORLD-X¹; BDH/ACS/ BDH1135 Methyl EthylOakwood/99.9%/075238 N/A N/A N/A 1147.8 kg 1149.4 kg Ketone (MEK) WaterMediatech/WFI/25-055-X¹; N/A N/A N/A   72.8 kg   72.5 kgRMBI/USP/WPW-USN-2XL Nitrogen Gas Airgas ≥ N/A N/A N/A Quantity Quantity(house system) 99% Sufficient Sufficient (QS)

-   1) CV8972 monohydrate (48.5 kg, CHP Lot #2479-1903-00489), Water    (48.5 kg, CHP Lot #329-190326), Methanol (19.2 kg, CHP Lot    #379-190404), and MEK (39.0 kg, CHP Lot #330-190326) were charged    respectively to a reactor R-402.-   2) The contents of the R-402 was adjusted to 20° C.±5° C. and    agitated until a solution was obtained.-   3) The contents of R-402 were transferred to R-401 through a 0.45    micron in-line filter.-   4) Water (24.0 kg, CHP Lot #329-190326) and Methanol (19.2 kg, CHP    Lot #379-190404) were charged to R-402 and transferred to R-401    through a 0.45 micron in-line filter.-   5) Methanol (192.1 kg, CHP Lot #379-190404) was charged to R-401    through a 0.45 micron in-line filter.-   6) The temperature of the contents of R-401 was adjusted to 60±5° C.-   7) MEK (899.6 kg, CHP Lot #330-190326; 380-190404) was charged to    R-401 through a 0.45 micron in-line filter over approx. 2 hours    while maintaining a temperature of 60±5° C.-   8) The contents of R-401 were agitated at 60±5° C. for at least 4    hours.-   9) The temperature of R-401 was adjusted to 20±5° C. over at least 3    hours.-   10) The contents of R-401 were agitated at 20±5° C. for approx. 9    hours.-   11) The contents of R-401 were sent to a filter.-   12) The filter cake was rinsed with MEK (105.4 kg, CHP Lot    #380-190404).-   13) The filter cake was rinsed with MEK (105.4 kg, CHP Lot    #380-190404).-   14) The wet cake were dried on filter with vacuum for at least 30    minutes.-   15) The wet cake was packaged into a filer dryer and dried under    reduced pressure at ≤30° C. for at least 12 hours.-   16) An IPC sample (IPC sample: 2479-1904-00494-32-01) was submitted    to QC for KF and Residual Solvent GC.    -   Water content (Specification: 2.8-3.8% by cKF)=3.5%    -   Residual Solvent GC Analysis (Specification: MeOH≤3000 ppm; MEK        ≤5000 ppm) MeOH=215 ppm; MEK=185 ppm-   17) The product CV-8972 was double bagged, goose necked, and    weighed.-   18) Analysis of the dried material (IPC sample:    2479-1904-00494-32-01)    -   Appearance: White to off-white solid    -   Weight=41.7 kg (86.0% yield)    -   HPLC purity (area %)=99.9%        -   Known Impurities:            -   Nicotinic Acid: No Peak            -   DMAP: No Peak            -   CV8814: 0.1%            -   2,3,4-Trimethoxy Benzaldehyde: No Peak            -   Trimetazidine: No Peak            -   CV-10099: No Peak            -   CV-10046: No Peak            -   XRPD: Conforms to Form A            -   Chloride Ion content: 19.4%            -   1H NMR: conforms to structure

Conclusion

The results provided above show that CV-8972 can be synthesized usingScheme 1. The reductive amination in step 1 using2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-ol startingmaterials, sodium triacetoxyborohydride (STAB) as the reductant,catalytic acetic acid (AcOH), and 2-methyltetrahydrofuran (2-MeTHF) gavea 78.0% yield of CV-8814 Free Base (CV8814 Free Base) with 100.0% purityby HPLC after aqueous workup, solvent exchange, and crystallization. A 5kg portion of CV-8814 Free Base (CV8814 Free Base) was diverted from thesynthesis for release. The step 2 coupling of CV8814 Free Base withnicotinic acid in the presence of EDC and catalytic DMAP in DCM went tocomplete conversion to CV8972 Free Base by HPLC IPC. Solvent exchange toMEK and addition into concentrated HCl in MEK afforded CV8972Monohydrate in a 96.4% yield with 99.1% purity by HPLC. The final formconversion in step 3 was completed by heating CV8972 Monohydrate to 60°C.±5° C. in a mixture of water, methanol, and MEK and precipitating outwith the addition of MEK. The white solid CV-8972 was obtained as form Aconfirmed by XRPD analysis, in an 86.0% yield with 99.9% purity by HPLC.The overall yield of the GMP synthesis of CV-8972 was 64.7%. The finalamount of CV-8972 produced was 41.7 kg.

INCORPORATION BY REFERENCE

References and citations to other documents, such as patents, patentapplications, patent publications, journals, books, papers, webcontents, have been made throughout this disclosure. All such documentsare hereby incorporated herein by reference in their entirety for allpurposes.

EQUIVALENTS

Various modifications of the invention and many further embodimentsthereof, in addition to those shown and described herein, will becomeapparent to those skilled in the art from the full contents of thisdocument, including references to the scientific and patent literaturecited herein. The subject matter herein contains important information,exemplification, and guidance that can be adapted to the practice ofthis invention in its various embodiments and equivalents thereof.

What is claimed is:
 1. A method for preparing a compound of Formula (X):

the method comprising the steps of: reacting2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-ol to producea free base form of a compound of Formula (IX):

and reacting the free base form of the compound of Formula (IX) withnicotinic acid to produce the compound of Formula (X), wherein themethod does not comprise producing a salt form of the compound ofFormula (IX).
 2. The method of claim 1, wherein the step of reacting2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-ol comprisessodium triacetoxyborohydride.
 3. The method of claim 1, wherein the stepof reacting 2,3,4-trimethoxybenzaldehyde and2-(piperazin-1-yl)ethan-1-ol comprises acetic acid.
 4. The method ofclaim 1, wherein the step of reacting 2,3,4-trimethoxybenzaldehyde and2-(piperazin-1-yl)ethan-1-ol comprises 2-methyltetrahydrofuran.
 5. Themethod of claim 1, wherein the step of reacting2,3,4-trimethoxybenzaldehyde and 2-(piperazin-1-yl)ethan-1-ol isperformed at from about 15° C. to about 25° C.
 6. The method of claim 1,wherein the step of reacting 2,3,4-trimethoxybenzaldehyde and2-(piperazin-1-yl)ethan-1-ol does not comprise dichloromethane.
 7. Themethod of claim 1, wherein the step of reacting the free base form ofthe compound of Formula (IX) with nicotinic acid produces a free baseform of the compound of Formula (X).
 8. The method of claim 7, whereinthe step of reacting the free base form of the compound of Formula (IX)with nicotinic acid comprises1-ethyl-3-(3-dimethylaminopropyl)carbodiimide.
 9. The method of claim 7,wherein the step of reacting the free base form of the compound ofFormula (IX) with nicotinic acid comprises 4-(dimethylamino)pyridine.10. The method of claim 7, wherein the step of reacting the free baseform of the compound of Formula (IX) with nicotinic acid comprisesdichloromethane.
 11. The method of claim 7, wherein the step of reactingthe free base form of the compound of Formula (IX) with nicotinic acidis performed at from about 20° C. to about 25° C.
 12. The method ofclaim 7, wherein the method further comprises converting the free baseform of the compound of Formula (X) to a salt form of the compound ofFormula (X).
 13. The method of claim 12, wherein the salt form of thecompound of Formula (X) is a HCl salt.
 14. The method of claim 12,wherein the salt form of the compound of Formula (X) is monohydrate. 15.The method of claim 12, wherein the step of converting the free baseform of the compound of Formula (X) to the salt form of the compound ofFormula (X) comprises HCl.
 16. The method of claim 12, wherein the stepof converting the free base form of the compound of Formula (X) to thesalt form of the compound of Formula (X) comprises methyl ethyl ketone.17. The method of claim 12, wherein the step of converting the free baseform of the compound of Formula (X) to the salt form of the compound ofFormula (X) is performed at about 50° C.
 18. The method of claim 12,wherein the method further comprises converting the salt form of thecompound of Formula (X) from a first crystal form to a second crystalform.
 19. The method of claim 18, wherein the step of converting thesalt form of the compound of Formula (X) from the first crystal form tothe second crystal form comprises one selected from the group consistingof precipitating the salt form of the compound of Formula (X), changingthe solvent of the salt form of the compound of Formula (X), andincubating the salt form of the compound of Formula (X), at about 60° C.20. The method of claim 1, wherein the method does not comprise use ofdioxane, ethylacetate, or potassium carbonate.
 21. A method forpreparing a compound of Formula (X):

the method comprising the steps of: reacting a compound of Formula (1):

with a compound of Formula (2):

to produce a free base form of a compound of Formula (IX):

reacting the free base form of a compound of Formula (IX) with acompound of Formula (3):

to produce a free base form of the compound of Formula (X); andconverting the free base form of the compound of Formula (X) to a HClsalt of the compound of Formula (X), wherein the method does notcomprise producing a salt form of the compound of Formula (IX).